Difference between revisions of "Part:BBa K1104101:Experience"

 
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This experience page is provided so that any user may enter their experience using this part.<BR>Please enter
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====Expression check====
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how you used this part and how it worked out.
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[[File:NYMU RFP-FimH(XP).png|right|thumb|300px|'''Check for the Length of the RFP-FimH''' <br>The electrophoresis gel shows the result after the digestion. The central well is the 100 bp marker.The two well on both sides of the marker is the digestion sample of RFP-FimH from different colony. The lower band is the part with the promoter and the RFP-FimH coding sequence, which has the length of 1,485 nt. The upper band is the backbone, pSB1A2, we use.]]
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[[File:NYMU_RFP-FimH cell.jpg|right|thumb|200px|'''Fluorescence Check''' <br>Both eppendorf are the liquid culture of the ''E. coli'' with RFP-FimH coding sequence cloned. We can obviously see the red light fluorescence emitted, showing that the fusion protein have successfully been produced and the RFP is still well-function.]]
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　　We ligated this part with LacI regulated promoter. In order to know whether this biobrick is correctly expressed in the ''E. coli'', we have done digestion on XbaI and PstI cutting site to proof that the length of our biobrick is right. So there is only the coding sequence of RFP-FimH instead of RFP only. The picture on the left is the result of the electrophoresis after the digestion. Secondly, through observing the fluorescence emitted by the ''E. coli'' we can be sure that we have produced the protein that we want, which is the RFP-FimH fusion protein. The result is as the picture on the right.<br>
  
 
===Applications of BBa_K1104101===
 
===Applications of BBa_K1104101===

Revision as of 20:08, 29 October 2013

Expression check

Check for the Length of the RFP-FimH
The electrophoresis gel shows the result after the digestion. The central well is the 100 bp marker.The two well on both sides of the marker is the digestion sample of RFP-FimH from different colony. The lower band is the part with the promoter and the RFP-FimH coding sequence, which has the length of 1,485 nt. The upper band is the backbone, pSB1A2, we use.
Fluorescence Check
Both eppendorf are the liquid culture of the E. coli with RFP-FimH coding sequence cloned. We can obviously see the red light fluorescence emitted, showing that the fusion protein have successfully been produced and the RFP is still well-function.

　　We ligated this part with LacI regulated promoter. In order to know whether this biobrick is correctly expressed in the E. coli, we have done digestion on XbaI and PstI cutting site to proof that the length of our biobrick is right. So there is only the coding sequence of RFP-FimH instead of RFP only. The picture on the left is the result of the electrophoresis after the digestion. Secondly, through observing the fluorescence emitted by the E. coli we can be sure that we have produced the protein that we want, which is the RFP-FimH fusion protein. The result is as the picture on the right.

Applications of BBa_K1104101

User Reviews

UNIQ45f2aa3e911fb791-partinfo-00000000-QINU UNIQ45f2aa3e911fb791-partinfo-00000001-QINU